Two-step resonance-energy-transfer-based ratiometric biosensor for sensing and annihilation of Staphylococcus aureus.

A two-step resonance energy transfer (RET)-based fluorescence/electrochemiluminescence (FL/ECL) biosensor was developed for ratiometric measurement and annihilation of Staphylococcus aureus (S. aureus). Using coupled dual-recognition-triggered target conversion with the catalytic hairpin assembly (CHA) technique, the monitoring of S. aureus was obtained at the single-cell level.

Ultrasensitive electrochemiluminescence biosensor based on polymethylene blue nanoparticles and DNA network for Staphylococcus aureus detection.

A novel bipolar electrode (BPE)-electrochemiluminescence (ECL) device was constructed for the ultra-sensitive detection of Staphylococcus aureus (S. aureus) by combining polymerase chain reaction (PCR) amplification and DNA network-loaded polymethylene blue nanoparticles (pMB NPs). The presence of target triggered the dissociation of double-stranded DNA on Fe3O4 NPs and the release of T strand, which initiated the PCR. The PCR product contains two protruding single-stranded DNA fragments that serve as bridges to connect Au NPs labeled probes. The PCR-Au products were captured by the probes on cathode of BPE to form three-dimensional DNA networks, which offer multiple adsorption sites for pMB NPs, leading to the remarkable enhancement of ECL intensity. Under optimal circumstances, a wide linear range from 10 to 108 CFU/mL and a low detection limit of 0.78 CFU/mL were achieved. This research opens new horizons for the application of PCR-based biosensors for the accurate and sensitive measurement of pathogenic bacteria.

Dual DNA recycling amplifications coupled with Au NPs@ZIF-MOF accelerator for enhanced electrochemical ratiometric sensing of pathogenic bacteria.

Sensitive and accurate quantification of pathogenic bacteria is vastly significant to the related food safety. Herein, a sensitive ratiometric electrochemical biosensor was developed for the detection of Staphylococcus aureus (S. aureus) based on dual DNA recycling amplifications and Au NPs@ZIF-MOF accelerator. Gold nanoparticles-loaded Zeolitic imidazolate metal-organic framework (Au NPs@ZIF-MOF) as electrode substrate possessed a large specific surface area for nucleic acid adsorption, and as an accelerator promoted the transfer of electrons. The strong recognition of aptamer to target S. aureus could initiate the padlock probe-based exponential rolling circle amplification (P-ERCA, as the first DNA recycling amplification), generating large numbers of trigger DNA strands. The released trigger DNA further activated the catalytic hairpin assembly (CHA, as the second DNA recycling amplification) on electrode surface. Consequently, P-ERCA and CHA continuously brought about one target to many signal transduction, leading to an exponential amplification. To achieve the accuracy of detection, the signal ratio of methylene blue (MB) and ferrocene (Fc) (IMB/IFc) was applied for intrinsic self-calibrating. Taking advantages of dual DNA recycling amplifications and Au NPs@ZIF-MOF, the proposed sensing system displayed high sensitivity for S. aureus quantification with a linear range of 5-108 CFU/mL, and the limit of detection was 1 CFU/mL. Moreover, this system represented excellent reproducibility, selectivity, and practicability for S. aureus analysis in foods.

Recent Advances in Electrochemiluminescence Biosensors for Mycotoxin Assay.

Rapid and efficient detection of mycotoxins is of great significance in the field of food safety. In this review, several traditional and commercial detection methods are introduced, such as high-performance liquid chromatography (HPLC), liquid chromatography/mass spectrometry (LC/MS), enzyme-linked immunosorbent assay (ELISA), test strips, etc. Electrochemiluminescence (ECL) biosensors have the advantages of high sensitivity and specificity. The use of ECL biosensors for mycotoxins detection has attracted great attention. According to the recognition mechanisms, ECL biosensors are mainly divided into antibody-based, aptamer-based, and molecular imprinting techniques. In this review, we focus on the recent effects towards the designation of diverse ECL biosensors in mycotoxins assay, mainly including their amplification strategies and working mechanism.

A dual-mode ratiometric aptasensor for accurate detection of pathogenic bacteria based on recycling of DNAzyme activation.

Pathogenic bacteria have a significant impact on food safety. Herein, an innovative dual-mode ratiometric aptasensor was constructed for ultrasensitive and accurate detection of Staphylococcus aureus (S. aureus) based on recycling of DNAzyme activation on gold nanoparticles-functionalized MXene nanomaterials (MXene@Au NPs). Electrochemiluminescent (ECL) emitter-labeled probe DNA (probe 2-Ru) containing the blocked DNAzyme was partly hybridized with aptamer and then captured by electrochemical (EC) indicator-labeled probe DNA (probe 1-MB) on electrode surface. When S. aureus presented, the conformation vibration of probe 2-Ru activated the blocked DNAzymes, leading to recycling cleavage of probe 1-MB and ECL tag close to electrode surface. Based on the reverse change tendencies of ECL and EC signals, aptasensor achieved S. aureus quantification from 5 to 108 CFU/mL. Moreover, the self-calibration characteristic of the aptasensor with dual-mode ratiometric readout ensured the reliable measurement of S. aureus in real samples. This work showed useful insight into sensing foodborne pathogenic bacteria.

Insights into the Mechanism of Bipolar Electrodeposition of Au Films and Its Application in Visual Detection of Prostate Specific Antigens.

Au particles are commonly used for deposition on the surface of a bipolar electrode (BPE) in order to amplify electrochemical and electrochemiluminescence (ECL) signal because of their excellent conductivity, biocompatibility, and large surface area. In this work, a closed BPE device was fabricated and Au particles were deposited on the two poles of a BPE via bipolar deposition. Results indicated that the electrochemical stability of Au film on the anode part of the BPE and the reduction of AuCl4- to Au on the cathode part of the BPE depended on the conductivity of the solution. The prepared Au-Au BPE exhibited a remarkable amplification effect on the ECL signal. Then, a specific sensing interface was constructed on one pole of the BPE for the visual detection of prostate-specific antigens (PSA) based on sandwich-type immunoreactions between primary PSA antibodies (Ab1) on the electrode surface, PSA, and SiO2 nanoparticles labeled secondary PSA antibodies (SiO2-Ab2). The designed biosensor exhibited a good linear relationship for the ECL detection of PSA in the range of 1 × 10-6 to 1 × 10-10 g/mL with a correlation coefficient of 0.9866; the limit of detection (LOD) was 1.5 × 10-11 g/mL. Additionally, the biosensor can realize the electrochemical imaging of PSA by regulating the electrochemical oxidation of the Au anode with the immunoreactions on the cathode part of BPE. Therefore, the small, portable and highly sensitive biosensors have great potential for on-site detection.

Novel electrochemiluminescence biosensor of fumonisin B1 detection using MWCNTs-PDMS flexible bipolar electrode.

A novel portable and disposable bipolar electrode (BPE)-electrochemiluminescence (ECL) device was fabricated for fumonisin B1 (FB1) detection. BPE was fabricated by using MWCNTs and polydimethylsiloxane (PDMS) due to their excellent electrical conductivity and good mechanical stiffness. After the deposition of Au NPs on the cathode of BPE, the ECL signal could be improved 89-fold. Then a specific aptamer-based sensing strategy was constructed by grafting capture DNA on Au surface, followed by hybridizing with aptamer. Meanwhile, an excellent catalyst, Ag NPs was labeled on aptamer to activate oxygen reduction reaction, leading to a 13.8-fold enhancement in ECL signal at the anode of BPE. Under the optimal conditions, the biosensor exhibited a wide linear range of 0.10 pg/mL to 10 ng/mL for FB1 detection. Meanwhile, it demonstrated satisfactory recoveries for real sample detection with good selectivity, making it to be a convenient and sensitive device for mycotoxin assay.

Rapid degradation of organic pollutants by Fe3O4@PDA/Ag catalyst in advanced oxidation process.

Over the past decades, the development of novel catalysts on the degradation of organic pollutants has attracted increasing attention. In this work, we synthesized silver decorated magnetic nanoparticles (Fe3O4@PDA/Ag NPs) to activate H2O2 for organic pollutants removal via advanced oxidation processes (AOPs). The catalyst was prepared through in-situ reduction of AgNO3 by the polydopamine (PDA) layer on Fe3O4 NPs. Chemiluminescence results obtained from luminol/H2O2 system revealed that the catalyst exhibited excellent catalytic effect on the decomposition of H2O2 into reactive oxygen species (ROS) and superoxide radical (O2-) was mainly responsible for the oxidative degradation. Importantly, the fast evolution frequency of oxygen gas bubbles produced in the reaction of Ag NPs and H2O2 could generate vigorous fluid convection and autonomous motion of catalyst when H2O2 concentration reached 1%. Additionally, the catalyst can suspend in solution for several minutes. Therefore, by coupling the vigorous motion with slow sedimentation velocity, the catalyst can realize rapid degradation of organic pollutants without external mixing force. The Fe3O4@PDA/Ag NPs catalysts not only showed a high removal efficiency of malachite green, but also can be applied for the degradation of other dyes, making it to be a promise candidate for environmental remediation. With the merits of excellent catalytic effect, fast degradation speed, and simplicity of operation, the prepared catalysts exhibits great potential in the practical field.

Novel integrating polymethylene blue nanoparticles with dumbbell hybridization chain reaction for electrochemical detection of pathogenic bacteria.

Pathogenic bacteria infections pose a major threat to human health which can be found in contaminated food and infected humans. Herein, an electrochemical sensor was developed for pathogenic bacteria assay using a dual amplification strategy of polymethylene blue nanoparticles (pMB NPs) and dumbbell hybridization chain reaction (DHCR). The strong binding ability of aptamer to targets endowed outstanding performance in identifying Staphylococcus aureus (S. aureus) among other typical bacteria. The released T strands were hybridized with capture DNA on electrode surface which triggered DHCR in the presence of two dumbbell-shaped helper DNA, leading to the formation of extended and tight dsDNA polymers. In combination with pMB NPs (redox indicators), S. aureus was quantitatively detected in a range of 10-108 CFU/mL and the detection limit reached 1 CFU/mL. Moreover, this sensor was successfully applied for S. aureus detection in human serum and foods, demonstrating the reliability in practical applications.

Soft-template solvent thermal method synthesis of magnetic mesoporous carbon-silica composite for adsorption of methyl orange from aqueous solution.

A facile soft-template solvent thermal strategy was developed to prepare mesoporous carbon-silica composite (MMCS) by using furfuryl alcohol (FA) as carbon precursor, Pluronic copolymer P123 as template, hydrated iron nitrate as iron source, and teraethylorthosilicate as silicon source and it was applied for the removal of methyl orange (MO). The as-synthesized MMCS with abound of hydrophilic groups processed a high specific surface area, large pore volume, and good magnetic response. With the increased amount of FA, the surface area and functional groups increased, promoting the adsorption effect. The maximum adsorption capacity of MO on MMCS can be high to 113.1 mg g-1 at pH 4 with 150 mg L-1 initial MO concentration. The adsorption isotherm, kinetic, and thermodynamics were all studied and the results showed the adsorption process well fitted Langmuir adsorption isotherm and pseudo-second-order model. Additionally, it was shown that the adsorption process could not be interfered by the co-existence of PO43-, NO3-, CO32-, SO42-, and real water matrix. And the proposed adsorbent can remove MO in three practical water samples with satisfied removal rates ranging from 92.8 to 99.8%. Thus, the MMCS prepared in this study could be utilized as an alternative adsorbent for the removal of methyl orange from practical aqueous solution.

Combination of area controllable sensing surface and bipolar electrode-electrochemiluminescence approach for the detection of tetracycline.

A novel area controllable biosensing interface is designed on glassy carbon bead (GCB) and used for measurement of tetracycline (TET) in closed bipolar electrode-electrochemiluminescence (ECL) device. One face of GCB is modified with Au particles and the covered area is varied from 0 to 45.3% by tuning the external voltage during bipolar electrodeposition process in a home-made open bipolar electrochemical cell. It enables the conjugation of various amounts of biomolecules on Au/GCB. Then DNA walker and methylene blue (MB) labeled DNA (MB-DNA) are conjugated on Au surface for the following combination of aptamer. In the presence of target, aptamer partially hybridized with DNA walker will be released from Au surface, leading to the formation of dsDNA between DNA walker and MB-DNA. As a result, MB-DNA with recognition site for nicking endonuclease (Nb.BbvCI) in dsDNA is cleft into two segments by Nb.BbvCI. Meanwhile, the liberated DNA walker is triggered to continue the degradation of many MB-DNA. Due to the excellent electrochemical performance of MB, it is reduced at the cathode of BPE to amplify the ECL signal at the anode of BPE in a closed BPE-ECL platform. When the covered area of GCB by Au particles is enlarged from 10.1 to 45.3%, the change of ECL intensity could be increased 2.7-fold. The linear range for the detection of tetracycline is from 1 × 10-12 to 1 × 10-5 M with the detection limit of 6.0 × 10-13 M. Hence, controllable sensing area on a single glassy carbon bead, the amplification effect of DNA walker and MB make this approach possess many advantages over traditional biosensor.

Bipolar electrode-electrochemiluminescence (ECL) biosensor based on a hybridization chain reaction.

A novel electrochemiluminescence (ECL) closed bipolar electrode (BPE) chip was designed based on a hybridization chain reaction (HCR)-induced ECL amplification strategy for the detection of both DNA and H2O2. Without the utilization of a patterned ITO bipolar electrode (BPE), this chip platform consisted of an ITO glass coated with two layers of PDMS slices. The ITO cathode was modified with Au nanoparticles for further functionalization of biomolecules, which could also amplify the ECL signal at the anode of the BPE. Based on the specific hybridization and hybridization chain reaction (HCR), DNA sequences were greatly extended, leading to a significant increase in the resistance of the cathode. The reduction of H2O2 was inhibited on the cathode of the BPE, resulting in a quenching effect on the ECL intensity on the anode of the BPE. The designed biosensor displayed a satisfactory linear relationship for the detection of both DNA and H2O2. Therefore, the biosensor could not only be employed for DNA assays but also used in enzyme reactions based on the generation of H2O2.

Buoyant force-induced continuous floating and sinking of Janus micromotors.

A novel bubble-induced ultrafast floating and sinking of micromotors based on the difference between buoyant force and gravity is proposed. Asymmetric micromotors were prepared by modification with Au and Pt layers for the two faces of glassy carbon beads (GCBs) by the bipolar electrodeposition technique. After the accumulation of enough oxygen bubbles by the decomposition of H2O2 at the Pt layer, the upward net force acting on the micromotor drove its movement to the air/solution interface. In order to reverse the direction of net force for the sinking of the micromotors, sodium dodecyl sulfate (SDS) was added into the fuel solution, which could facilitate the release of bubbles and decrease the diameter of the bubbles. However, the lifetime of the bubbles was increased significantly. After the addition of a small amount of salt, the lifetime of the bubbles was obviously reduced. As a consequence, the breakup of bubbles on the micromotor changed the direction of the net force from up to down which pulled the micromotor down to the bottom of the solution. The velocity of the micromotor was dependent on the net force exerted on the micromotor, leading to an ultrafast motion of the micromotor. It still reached 1.2 cm s-1 after 3 h. Moreover, the simple asymmetric deposition technique showed great promise for the further application of the micromotors in bioanalysis and environmental remediation.

Roles of L-type calcium channels (CaV1.2) and the distal C-terminus (DCT) in differentiation and mineralization of rat dental apical papilla stem cells (rSCAPs).

OBJECTIVE: Voltage-gated inward Ca2+ currents (ICa) are triggered by cell depolarization and commonly produce transient increases in the cytoplasmic free Ca2+ concentration. The CaV1.2 distal C-terminus is susceptible to proteolytic cleavage, which yields a truncated CaV1.2 subunit and a cleaved C-terminal fragment (CCt or DCT). Stem cells from the apical papilla (SCAPs) has a capacity for differentiation into the odontoblastic-like cells in vitro and dentin forming in vivo, which makes SCAPs advantages in tissue engineering and regenerative endodontic. The aim of this study was to investigate the effect of CaV1.2 and its distal C-terminal fragment in the odontoblastic differentiation of rat SCAPs (stem cells from the apical papilla). DESIGN: In this study, we generated stable CaV1.2 knockdown and DCT over-expressed rSCAPs using short hairpin RNA and DCT gene containing Lentivirus vectors, respectively. The transfected apical papilla cells were induced to differentiate into the odontoblast-like cells, and the expression of markers for odontoblastic differentiation were analyzed by alizarin red staining, Real-time Polymerase chain reaction (RT-PCR), and Western blot analysis. RESULTS: The knockdown of CaV1.2 and excess expression of DCT both suppressed the expression of DSPP, ALP in mRNA level and the formation of calcium nodules. CONCLUSIONS: Our results suggest that CaV1.2 and DCT play important roles in the differentiation of rSCAPs, DCT might act as a transcription factor and regulate the differentiation of rSCAPs.

Novel "signal-on" electrochemiluminescence biosensor for the detection of PSA based on resonance energy transfer.

Here, we propose a simple and novel "signal-on" electrochemiluminescence (ECL) biosensor based on resonance energy transfer (RET) for detection of prostate specific antigen (PSA). The system is composed of Multi-walled carbon nanotubes (MWCNT), polyamidoamine (PAMAM) dendrimer and Au nanoparticles (NPs) film on glassy carbon electrode (GCE) to improve the electron transfer, provide abundant amine group for the immobilization of biomolecules, and amplify the ECL signal. After that, Au nanorods (Au NRs) labeled peptide is modified on electrode surface to serve as ECL-RET acceptor due to the excellent overlap between the ECL emission spectrum of Ru(bpy)32+ and the absorption spectrum of Au NRs, leading to the significant decrement of ECL signal. Upon the sensing cleavage of peptide with PSA, both Au NRs and peptide are released from electrode surface, resulting in the high recovery efficiency of ECL signal. The proposed approach exhibits a wide linear range from 0.1pg/mL to 10ng/mL with a detection limit of 0.03pg/mL. Results revealed that the recoveries were in a range from 95% to 108%, indicating good accuracy of the proposed method for PSA detection. In addition, the proposed biosensor exhibited well specificity for the detection of PSA.

Joint enhancement strategy applied in ECL biosensor based on closed bipolar electrodes for the detection of PSA.

A highly sensitive electrogenerated chemiluminescence (ECL) biosensor was developed on the basis of a closed bipolar electrode (BPE) apparatus for the analysis of prostate specific antigen (PSA). Bipolar modifications bring up two different stages of enhancement on the same electrode. Anodic enhancement was conducted by modifying gold nanoparticles (Au NPs) to catalyze the anodic ECL reaction between luminol and hydrogen peroxide. Cathodic introduction of thionine tagged PSA antibody led to a further pertinently enhancement synchronized with the PSA amount variation, because the existence of thionine greatly increased the rate of electron gains on cathode, leading to the corresponding acceleration of anodic ECL reaction. The more thionine modified target molecules were introduced, the faster luminol was oxidized, the higher faraday current approached, and sensitive quantification was realized in correlation with the responsive ECL intensity differences. The quantification resulted in a good determination range between 0.1pg/mL and 0.1µg/mL. This strategy mainly took advantage of the special structure of closed BPE to realize a simultaneous amplification on both ends of BPE. Moreover, the platform had a potential of providing a multi-functional strategy for the realization of other bio-detections by simply substituting the PSA sandwich structure with other bio-structures.

Effects of cold light bleaching on the color stability of composite resins.

To evaluate the effects of cold light bleaching on the color stability of four restorations using a thermocycling stain challenge. 160 specimens (10 mm in diameter and 2 mm thick) were fabricated from 4 composite resins (Gradia Direct-A, Z350XT, Premisa, and Précis) and divided into 4 subgroups. Color was assessed according to the CIEL*a*b* color scale at baseline, after the first cycle of bleaching, after thermocycling stain challenges, and after the second cycle of bleaching. Mean values were compared using three-way analysis of variance, and multiple comparisons of the mean values were performed using the Tukey-Kramer test. All groups showed significant color changes after stain challenge, the color change was more significant in Gradia Direct and Z350XT than in Premisa and Précis. After the second cycle of bleaching, color mostly recovered to its original values. The color stability of Gradia Direct and Z350XT was inferior to that of Premisa and Précis. The discoloration of composite resin materials can be partly removed after cold light bleaching.

Mesoporous silica film-assisted amplified electrochemiluminescence for cancer cell detection.

A mesoporous silica film-assisted amplification method is reported for the first time for the sensitive electrochemiluminescence detection of cancer cells.

Visual electrochemiluminescence detection of cancer biomarkers on a closed bipolar electrode array chip.

This paper describes a novel electrochemiluminescence (ECL) imaging platform for simultaneous detection of cancer biomarkers based on a closed bipolar electrode (BPE) array. It consists of two separated channel arrays: detection channel array and sensing channel array, which are connected by a group of parallel ITO BPEs on a glass substrate. Besides, two parallel ITO strips are fabricated at the two sides of BPE array and employed as driving electrodes. After Au films are electrochemically deposited on the cathodes of the BPE array, nanobioprobes including biorecognition elements (aptamer or antibody) and a novel electrochemical tag, which is synthesized by doping thionine in silica nanoparticles (Th@SiO2 NPs), are introduced into the cathodes by immunoreaction or DNA hybridization. The Th@SiO2 coupled nanobioprobes as both recognition probes and signal amplification indicators could mediate the ECL signals of Ru(bpy)3(2+)/tripropylamine (TPA) on the anodes of BPE array through faradaic reaction due to the charge neutrality of BPE. Thus, multiplex detection of cancer biomarkers (adenosine triphosphate (ATP), prostate-specific antigen (PSA), α-fetoprotein (AFP) and thrombin) is realized by forming specific sensing interfaces onto the cathodic poles of BPEs in different sensing channels and reported by the ECL images of the Ru(bpy)3(2+)/TPA system on the anodic poles of BPEs in detection channels. The results demonstrate that this visual ECL platform enables sensitive detection with excellent reproducibility, which may open a new door toward the development of simple, sensitive, cost-effective, and high throughput detection methods on biochips.

Visual electrochemiluminescence detection of telomerase activity based on multifunctional Au nanoparticles modified with G-quadruplex deoxyribozyme and luminol.

A novel visual electrochemiluminescence (ECL) analysis strategy for detection of telomerase activity is reported on a microarray chip, with G-quadruplex deoxyribozyme (DNAzyme) and luminol modified Au nanoparticles (NPs) as double-catalytic amplification labels.